Choeradoplana Crassiphalla Sp. Nov. (Platyhelminthes: Tricladida: Geoplanidae): a New Species of Land Planarian from the Atlanti

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Studies on Neotropical Fauna and Environment Publication details, including instructions for authors and subscription information: http://www.tandfonline.com/loi/nnfe20 Choeradoplana crassiphalla sp. nov. (Platyhelminthes: Tricladida: Geoplanidae): a new species of land planarian from the Atlantic Forest of Argentina Lisandro Negrete a b & Francisco Brusa a b a División Zoología Invertebrados, Facultad de Ciencias Naturales y Museo, Universidad Nacional de La Plata, La Plata, Argentina b CONICET Version of record first published: 30 Nov 2012.

To cite this article: Lisandro Negrete & Francisco Brusa (2012): Choeradoplana crassiphalla sp. nov. (Platyhelminthes: Tricladida: Geoplanidae): a new species of land planarian from the Atlantic Forest of Argentina, Studies on Neotropical Fauna and Environment, 47:3, 227-237 To link to this article: http://dx.doi.org/10.1080/01650521.2012.735071

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Choeradoplana crassiphalla sp. nov. (Platyhelminthes: Tricladida: Geoplanidae): a new species of land planarian from the Atlantic Forest of Argentina Lisandro Negretea,b & Francisco Brusaa,b* aDivisión Zoología Invertebrados, Facultad de Ciencias Naturales y Museo, Universidad Nacional de La Plata, La Plata, Argentina; bCONICET (Received 25 January 2012; accepted 26 September 2012)

A new species of land planarian from northeastern Argentina, Choeradoplana crassiphalla sp. nov., is described. This is the ﬁrst report of Choeradoplana in the Atlantic Forest of Argentina and the second species of the genus for this country. Choeradoplana includes nine species of Neotropical land planarians. The new species is light brown with a thin clearer median stripe. The pharynx is bell-shaped. The copulatory apparatus presents a short and very high and thick penis papilla. Choeradoplana crassiphalla is compared with other species of the genus, especially with Ch. marthae, due to the similarity of the copulatory apparatus. Keywords: terrestrial ﬂatworm; Geoplaninae; Choeradoplana species; northeastern Argentina

Introduction There are no records of land planarian species in The genus Choeradoplana includes nine species of the Argentinean portion of the Atlantic Forest. Only Neotropical land planarians. Eight of the nine known Bipalium kewense Moseley, 1878 was found in the species have been described for Brazil (von Graff vicinity of native forests (Negrete et al. 2011). The 1899; Marcus 1951; Froehlich 1955; Carbayo & Atlantic Forest of South America is one of the world’s Froehlich 2012), whereas Ch. langi (von Graff, 1894) 25 recognized biodiversity hotspots, with more than has been described for Paraguay, and recorded in 60% of all terrestrial species on earth (Galindo-Leal & Argentina (Buenos Aires province) and Brazil (Du Câmara 2003). The Atlantic Forest is a complex of Bois-Reymond Marcus 1951; Ogren & Kawakatsu ecoregions that span the Atlantic coast of Brazil, 1990). These species are characterized by their small extending westward into eastern Paraguay and north- size (20–50 mm length) and the anterior tip dilated eastern Argentina (Misiones province) (Di Bitetti et al. and rolled towards the back when the flatworms crawl 2003). above the soil. The anterior tip possesses two glan- The aim of this paper is to describe a new species dular cushions, separated by a median groove. The of land planarian of the genus Choeradoplana from the glandular cushions are the glandulo-muscular organ, Atlantic Forest of Argentina. named “Drüsenpolster” by von Graff (1899). Also, the genus is characterized by possessing part of the cuta- Downloaded by [Duke University Libraries] at 08:39 12 December 2012 neous longitudinal muscle layer partially sunk into the Material and methods parenchyma. The cephalic retractor muscle (derived The land flatworms were collected between 2009 and from that layer) has a triangular shape in cross section 2010 in the Reserva de Vida Silvestre Urugua-í ◦ ◦ (Carbayo & Leal-Zanchet 2003). (RVSU)(25 59 S, 54 05 W) and Campo Anexo INTA ◦ ◦ The land planarian fauna from Argentina is poorly in San Antonio County (SA) (26 02 S, 53 47 W), known. Most of the descriptions and records date Misiones province, Argentina. The two sampling loca- back to the nineteenth century (von Graff 1894, 1897, tions belong to the Atlantic Forest ecosystem. The first 1899). With the recent record of a non-native species, locality is within the Upper Paraná Atlantic Forest Caenoplana coerulea Moseley, 1877 (Negrete et al. ecoregion, characterized by a semideciduous seasonal 2011), 14 species of land planarians (Geoplanidae) forest, with trees ranging in height from 20 to 50 m, are known for Argentina. Three subfamilies are rep- with strata of smaller trees and a dense undergrowth resented: 10 species of Geoplaninae, three species of of bamboo (Giraudo et al. 2003). The second one Rhynchodeminae and one species of Bipaliinae. belongs to the Araucaria moist forest ecoregion, with

*Corresponding author. Email: [email protected]

the predominance of the Paraná pine Araucaria angus- anterior region 2: sagittal sections on 14 slides; pre- tifolia and tree ferns. pharyngeal region: transverse sections on four slides; The geoplanids were manually collected during the pharynx and copulatory apparatus: sagittal sections day beneath fallen logs, leaf litter and stones, and on 19 slides. MLP 6562. Argentina, Misiones, RVSU, during the night, when planarians are active. The ani- 21 August 2009, L. Negrete, coll.; anterior region: mals were photographed in vivo and their external transverse sections on eight slides; pre-pharyngeal morphology and color pattern were recorded. The region: transverse sections on three slides; pharynx specimens were killed with boiling water, ﬁxed in 10% and copulatory apparatus: sagittal sections on nine formaldehyde and conserved in 70% ethanol. Body slides. fragments were dehydrated in an ascending series of ethanol and embedded in Paraplast®.Sagittaland transverse serial sections (8 µm thick) of the anterior Additional material examined body region, transverse sections of the pre-pharyngeal MLP 6563. Argentina, Misiones, RVSU, 22 August region (8 µm thick) and sagittal serial sections of the 2009, L. Negrete, coll.; preserved in 70% ethanol. pharynx and the copulatory apparatus (8 µm thick) MLP 6564. Argentina, Misiones, SA, 18 May 2009, were stained with Masson’s trichrome method and L. Negrete, coll.; preserved in 70% ethanol. hematoxylin–eosin method (Romeis 1989). The ratio of the height of cutaneous musculature to the height Diagnosis of the body (cutaneous muscular index, CMI) was calculated according to Froehlich (1955). The copula- The new species can be distinguished from the other tory apparatus of the specimens was reconstructed for species in the genus by the following particular com- descriptive and diagnostic purposes. bination of features: light brown colored dorsal sur- The holotype, paratypes and additional specimens face with a thin clear median longitudinal stripe, and were deposited in the Invertebrate Collection at Museo yellowish brown body margin; intra-bulbar prostatic de La Plata (MLP), Argentina. vesicle consisting of a large and highly folded cham- ber; the short, high and thick penis papilla – ventrally to the prostatic vesicle – hanging from the roof of the male atrium; female atrium 3–5 times shorter than Results male atrium. Family Geoplanidae Stimpson, 1857 Subfamily Geoplaninae Stimpson, 1857 Genus Choeradoplana von Graff, 1896 Etymology Choeradoplana crassiphalla sp. nov. The speciﬁc name is derived from the Latin: crassus (Figures 1–7) (thick) and phallus (penis), due to the features of the penis papilla.

Type series Type locality Holotype: MLP 6561. Argentina, Misiones, RVSU, 23 May 2009, L. Negrete, coll.; anterior region: trans- Reserva de Vida Silvestre Urugua-í, northern Misiones province, Argentina. Downloaded by [Duke University Libraries] at 08:39 12 December 2012 verse sections on five slides; pre-pharyngeal region: transverse sections on five slides; pharynx and copulatory apparatus: sagittal sections on 11 slides. Description Paratypes: MLP 6225. Argentina, Misiones, SA, 16 May 2009, L. Negrete, coll.; anterior region: sagittal External morphology sections on 23 slides; pre-pharyngeal region: trans- Dorsum brown, ranging from light to darker pigmen- verse sections on 14 slides; pharynx and copula- tation. Very thin median longitudinal stripe, lighter tory apparatus: sagittal sections on 15 slides. MLP than ground color, being 5% with respect to body 6226-3. Argentina, Misiones, RVSU, 22 August 2009, width (Figure 1A, B). Median stripe starting posteri- L. Negrete, coll.; anterior region 1: transverse sec- orly of light gray anterior tip, continuing throughout tions on 12 slides; anterior region 2: sagittal sections the body and disappearing just before the posterior on eight slides; pre-pharyngeal region: transverse sec- end. Body margins yellowish brown, ventral surface tions on five slides; pharynx and copulatory apparatus: pale yellowish brown to whitish. Some specimens with sagittal sections on 12 slides. MLP 6481. Argentina, posterior end black. Anterior tip blunt, ventrally with Misiones, SA, 5 November 2010, L. Negrete, coll.; striations separated by a median groove. Posterior end anterior region 1: transverse sections on 15 slides; narrowing gradually. Studies on Neotropical Fauna and Environment 229

Figure 1. Photographs of living specimens of Choeradoplana crassiphalla sp. nov. (A) Holotype, with harvestman trapped in mucus. Scale bar: 10 mm. (B) Paratype MLP 6226-3, at rest. Scale bar: 10 mm. C. Detail of the anterior end of another specimen. Scale bar: 5 mm.

On both sides eyes beginning 1–2 mm behind secretion, and a retractor muscle. Secretions form-

Downloaded by [Duke University Libraries] at 08:39 12 December 2012 anterior end, being absent at the tip. Eyes initially uni- ing two glandular cushions, separated by a median serial and marginal, then bi- or triserial and marginal, groove, on both sides of retractor muscle (Figure 2A). very close to each other for 4–5 mm (Figure 1C). Fibers of retractor muscle concentrated in median Thereafter, eyes bi- or uniserial, some scattered, less ventral region, beneath nervous plate. Retractor mus- numerous at mouth level, reaching posterior end of the cle triangle-shaped in cross section (Figure 2A). Thin body. layer of transversal muscles (5 µm thick) present After ﬁxation, body length between 22 and between nervous plate and retractor muscle, thus 43 mm, maximum width 1.5–3.5 mm, maximum forming a sub-neural layer. In anterior region, supra- height 0.9–1.9 mm. Mouth located at a distance intestinal ﬁbers well developed forming an inter- of 55.5–62.5% from anterior tip; gonopore at mingled muscle (Figure 2A). Fibers of cutaneous 66.3–73.3%, respectively (Table 1). longitudinal muscle layer partially sunk into the parenchyma. Sensory pits as simple invaginations (30–40 µm Internal morphology deep) present in ventral body margins (Figure 2B). Cephalic region with musculo-glandular organ, con- They appear posteriorly from anterior tip, forming a sisting of very abundant erythrophil and rhabditogen sensory border until 1–1.5 mm from anterior tip. 230 L. Negrete & F. Brusa Downloaded by [Duke University Libraries] at 08:39 12 December 2012

Figure 2. Transverse sections of the cephalic region of the body of Choeradoplana crassiphalla sp. nov. (paratype MLP 6481). (A) Close to the anterior tip. (B) Immediately behind the glandular cushions. Abbreviations: dp: dorsal parenchymatic muscle layer; e: eye; i: intestine; lc: longitudinal cutaneous muscle layer; n: nervous plate; r: retractor muscle; rb: rhabdites; rbb: rhabditogen cellular bodies; s: sensory pit; sb: sub-intestinal transverse parenchymatic muscle layer; sn: sub-neural transverse parenchymatic muscle layer; sp: supra-intestinal transverse parenchymatic muscle layer. Scale bars: 250 µm. Studies on Neotropical Fauna and Environment 231

Figure 3. Sagittal section of the pharynx of Choeradoplana crassiphalla sp. nov. (holotype). Abbreviations: di: dorsal insertion of the pharynx; i: intestine; mo: mouth; phg: pharyngeal glands; ph: pharyngeal pouch; pl: pharyngeal lumen; t: testes; vi: ventral insertion of the pharynx. Scale bar: 500 µm. Downloaded by [Duke University Libraries] at 08:39 12 December 2012

Figure 4. (A) Detail of a sagittal section of the anterior region of the body at the level of the ovaries of Choeradoplana crassiphalla sp. nov. (paratype MLP 6481). (B) Detail of a sagittal section of the pre-pharyngeal region of Choeradoplana crassiphalla sp. nov. (paratype MLP 6481). Abbreviations: dc: diagonal cutaneous muscle layer; dp: dorsal parenchymatic muscle layer; dv: dorsoventral parenchymatic muscles; i: intestine; lc: longitudinal cutaneous muscle layer; n: nervous plate; od: ovovitelline duct; ov: ovary; sb: sub-intestinal transverse parenchymatic muscle layer; sp: supra-intestinal transverse parenchymatic muscle layer; t: testes; vit: vitellaria. Scale bars: 100 µm (A), 200 µm(B).

Ventral epidermis ciliated on creeping sole, which margins (Figure 2B). In ventral epidermis secretion is approximately 70–85% of body width at pre- is less abundant, occupying the apex of the cells. pharyngeal region. Ventral epidermis 20–35 µm high; Rhabditogen cellular bodies located beneath cuta- dorsal epidermis 15–20 µm high. Rhabdites very neous longitudinal muscle layer, or among ﬁbers of abundant over entire dorsal epidermis and body dorsal parenchymatic muscle layer. Scarce cyanophil 232 L. Negrete & F. Brusa

Figure 5. (A) Sagittal section of the copulatory apparatus of Choeradoplana crassiphalla sp. nov. (paratype MLP 6226-3). (B) Detail of a sagittal section of Choeradoplana crassiphalla sp. nov. (holotype), showing the prostatic vesicle. Abbreviations: cm: common muscle coat; ed: efferent duct; ej: ejaculatory duct; i: intestine; ma: male atrium; od: ovovitelline duct; pb: penis bulb; pg: prostatic glands; pp: penis papilla; pv: prostatic vesicle; sg: shell glands. Scale bars: 500 µm.

secretion discharging through epidermis. Glandular beneath supra-intestinal parenchymatic muscle layer body margin absent. (Figure 4A). Testes arranged in two or three rows on Cutaneous musculature composed of an external each side of body. Foremost testes anterior of ovaries, layer of circular muscles, a diagonal layer, and an inter- spreading until close to pharynx (Figure 3, Table 3). nal longitudinal layer formed by bundles (Figure 2B, Testes height between 10.5% and 25% of body Table 2). The latter most strongly developed and ven- height. trally subdivided into two layers. Innermost layer sunk Efferent ducts at pre-pharyngeal level, run- into parenchyma, located between cutaneous nerve net ning above and among fibers of sub-intestinal and nervous plate. The CMI ranging from 16.3% to parenchymatic muscle layer, located slightly dorsal to 28.3% (Table 2). ovovitelline ducts. Efferent ducts lined with a cili- Parenchymatic musculature arranged in three lay- ated squamous epithelium. Distally, behind pharynx, ers with similar thickness: a dorsal layer with diag- efferent ducts widened like false seminal vesicles, onal fibers, a supra-intestinal and a sub-intestinal their lumen full of spermatozoa (Figure 5A), and transverse layer (Figure 2B). Also, dorsoventral their epithelium cuboidal. Distal tracts of efferent fibers intermingled among the intestinal branches. ducts entering penis bulb, bending almost 90◦ towards The thickness of the parenchymatic musculature dorsum and connecting laterally with first third of represents 34.2–43% of the cutaneous musculature prostatic vesicle (Figures 5A, 7). Prostatic vesicle (Table 2). located inside the common muscle coat, therefore Pharynx bell-shaped. Mouth located in the middle being intra-bulbar (Figures 5B, 6, 7), dorsal of male

Downloaded by [Duke University Libraries] at 08:39 12 December 2012 of pharyngeal pouch (Figure 3). Pharynx 1.27–1.7 mm atrium, oval shaped and its walls folded (Figures 5B, long. Pharyngeal epithelium cuboidal and ciliated. 6B, 7). Its length variable (Table 3). Prostatic vesi- Inner musculature of pharynx well developed, being cle lined with ciliated cylindrical epithelium, pro- composed of a circular subepithelial layer (15–30 µm vided with abundant xanthophil granulous secretion. thick), under which there is a longitudinal somewhat Secretion highly abundant, being discharged from thicker layer (15–50 µm thick). External muscula- secretory cells whose cellular bodies are located ture of pharynx less developed than internal, being both anterior and posterior to prostatic vesicle, and arranged in circular (25–30 µm thick) and longitu- both inside and outside of common muscle coat dinal (5–20 µm thick) ﬁbers, respectively. Numerous (Figures 6, 7). Prostatic vesicle joining ejaculatory cell necks of erythrophil secretory cells and, less abun- duct, which runs almost concentrically along the penis dant, cyanophil cell necks, whose cellular bodies are papilla. Ejaculatory duct with cuboidal epithelium and found in the parenchyma of the pre-pharyngeal region, receiving scarce cyanophil and xanthophil secretions traversing the pharynx to open through the pharyngeal (Figure 7). epithelium (Figure 3). Penis papilla short, very high and thick Testes of all specimens mature, with spermatozoa (Figures 6, 7, Table 3), hanging from the roof of in their lumen. Testes spherical or oval, dorsal, located the male atrium, occupying most of its cavity. Penis Studies on Neotropical Fauna and Environment 233 Downloaded by [Duke University Libraries] at 08:39 12 December 2012

Figure 6. Sagittal section of the copulatory apparatus of (A) holotype and (B) paratype MLP 6226-3 of Choeradoplana crassiphalla sp. nov. Abbreviations: cm: common muscle coat; cod: common ovovitelline glandular duct; ej: ejaculatory duct; fa: female atrium; g: gonopore; i: intestine; ma: male atrium; od: ovovitelline duct; pg: prostatic glands; pp: penis papilla; pv: prostatic vesicle; sg: shell glands; v: vagina; vit: vitellaria. Scale bars: 500 µm.

papilla dorsoventrally to slightly obliquely oriented female (Figures 6, 7, Table 3). Its epithelium rang- (Figures 6, 7), with cuboidal epithelium receiving ing between cuboidal and cylindrical, apically abundant erythrophil and scarce cyanophil secre- erythrophil. Muscularis of male atrium composed of tions. Its stroma possessing abundant intermingled a subepithelial longitudinal muscle layer (5–12 µm muscle ﬁbers. Male atrium 3–5 times longer than thick) followed by a circular one (12–17 µm thick). 234 L. Negrete & F. Brusa Downloaded by [Duke University Libraries] at 08:39 12 December 2012

Figure 7. Diagrammatic sagittal reconstruction of the copulatory apparatus of (A) holotype and (B) paratype MLP 6226-3 of Choeradoplana crassiphalla sp. nov. Abbreviations: cm: common muscle coat; cod: common ovovitelline glandular duct; ed: efferent duct; ej: ejaculatory duct; fa: female atrium; g: gonopore; ma: male atrium; od: ovovitelline duct; pb: penis bulb; pg: prostatic glands; pp: penis papilla; pv: prostatic vesicle; sg: shell glands; v: vagina. Scale bars: 1 mm. Studies on Neotropical Fauna and Environment 235

Table 1. Measurements (mm) of the specimens of Choeradoplana crassiphalla sp. nov. All the measurements were obtained from ﬁxed specimens.

Holotype Paratype Paratype Paratype Paratype MLP 6561 MLP 6226-3 MLP 6562 MLP 6481 MLP 6225

Length 22 43 24 43 38 Maximum width 1.5 3.5 1.8 2.15 2.2 Height 0.95 1.4 0.9 1.4 1.93 DM 12.9 26.5 15 26 21.1 DM (%) 58.6 61.6 62.5 60.5 55.5 DG 15.7 31.5 17 31 25.2 DG (%) 71.4 73.3 70.8 72.1 66.3 DMG 2.8 5 2 5 4.1 CS (%) 75 71.3 70 76 76.4

Note: Abbreviations: DG: distance of gonopore from anterior end; DG (%): DG/body length; DM: distance of mouth from anterior end; DM (%): DM/body length. DMG: distance between mouth and gonopore; CS: creeping sole width.

Table 2. Thickness of cutaneous (CM) and parenchymatic (PM) musculature (µm) and CMI in the pre-pharyngeal region of specimens of Choeradoplana crassiphalla sp. nov.

Holotype Paratype Paratype Paratype Paratype MLP 6561 MLP 6226-3 MLP 6562 MLP 6481 MLP 6225

CM dorsal circular 5 2.5 2.5 5 5 CM dorsal diagonal 20 12.5 15 40 40 CM dorsal longitudinal 100 95 87.5 145 120 CM dorsal total 125 110 105 190 165 CMventralcircular55555 CM ventral diagonal 15 12.5 20 25 20 CM ventral longitudinal 100 175 125 175 125 CM ventral total 120 192.5 150 205 150 CMI (%) 25.8 21.6 28.3 18.4 16.3 PMdorsal2550355050 PM supraintestinal 37.5 50 25 60 50 PM subintestinal 25 30 30 25 35 PM:CM total (%) 35.7 43 35.3 34.2 42.9

Table 3. Measurements (mm) of the reproductive system of specimens of Choeradoplana crassiphalla sp. nov.

Holotype Paratype Paratype Paratype Paratype MLP 6561 MLP 6226-3 MLP 6562 MLP 6481 MLP 6225

Anteriormost testes 4.8 (21.8%) – – 4 (9.3%) 8.25 (21.7%) Posteriormost testes 11.8 (53.6%) 25 (58.1%) 13.9 (57.9%) 23.5 (54.6%) 19.6 (51.6%)

Downloaded by [Duke University Libraries] at 08:39 12 December 2012 T:BH 23% 18.7% 25% 10.5% 15.5% LPV × HPV 0.65 × 0.21 1.25 × 0.5 0.65 × 0.3 0.62 × 0.28 0.8 × 0.35 LPP × WPP × HPP 0.3 × 0.7 × 10.3× 0.92 × 1.3 0.25 × 0.7 × 0.6 0.15 × 0.6 × 0.85 0.3 × 0.42 × 1.1 LMA 0.85 1.32 0.55 0.88 1.2 Location of ovaries 4.9 (22.3%) – – 4.1 (9.5%) 8.5 (22.3%) LGCD 0.09 0.08 0.15 0.08 0.12 LV 0.13 0.38 0.28 0.13 0.32 LFA 0.3 0.25 0.2 0.3 0.25 LFA:LMA 35.3% 18.9% 36.4% 34.1% 20.8%

Note: Abbreviations: LFA: length of female atrium; LGCD: length of common glandular duct; LMA: length of male atrium; LPP: length of penis papilla; WPP: width of penis papilla; HPP: height of penis papilla; LPV: length of prostatic vesicle; HPV: height of prostatic vesicle; LV: length of vagina; T: testes height; BH: body height.

Erythrophil secretion discharging through epithelium (Figure 4B). Ovaries situated at a distance of ∼20% of male atrium. body length from anterior end. In one young specimen Ovaries located ventrally, between sub-intestinal (without shell glands) ovaries situated at a distance parenchymatic muscle layer and nervous plate of 9.5% of body length (Table 3). Ovaries with 236 L. Negrete & F. Brusa

globose anterior portion and long, narrow posterior Carbayo & Froehlich, 2012, Ch. catua Froehlich, 1955, one (Figure 4B). Ovovitelline ducts emerging lat- Ch. gladismariae Carbayo & Froehlich, 2012, Ch. erally and externally from first portion of ovaries. iheringi von Graff, 1899, and Ch. langi, the male repro- In several specimens, proximal part of ovovitelline ductive system has a folded male atrium. However, in ducts with spermatozoa. In the pre-pharyngeal Ch. catua and Ch. langi the atrium could act as a tem- region, ovovitelline ducts immediately beneath porary penis papilla. The species that have a folded sub-intestinal parenchymatic muscle layer. Their male atrium have a prostatic vesicle different from the epithelium cuboidal and ciliated. Ovovitelline ducts new species. This prostatic vesicle is anterior to the running backwards, reaching copulatory apparatus, copulatory apparatus; in the new species it is dorsal and then taking an ascending direction immediately to the copulatory apparatus. behind gonopore, bending toward sagittal plane and In Ch. bilix Marcus, 1951 and Ch. marthae opening into common glandular ovovitelline duct Froehlich, 1955, there is a penis papilla like in Ch. (Figure 7). Distal ascending portions of ovovitelline crassiphalla sp. nov. The new species can be exter- ducts and of the common glandular duct receiving nally distinguished from Ch. bilix due to the latter abundant secretion from shell glands (Figure 7). having a yellow ground color at the dorsum with Short common glandular duct lined with ciliated two lateral grayish brown bands. Regarding the inter- cuboidal high epithelium. This is dorsoventral and nal morphology, the pharynx of Ch. bilix is between slightly curved forward, and in some specimens is bell-shaped and the collar type, and the prostatic C-shaped. Common glandular duct opening into vesicle is extra-bulbar and anterior to the penis a long vagina (Figures 6A, 7, Table 3). Epithelium papilla. of vagina ranging from non-ciliated cuboidal high Choeradoplana crassiphalla sp. nov. resembles Ch. to cylindrical. Muscularis of vagina composed of a marthae, mainly in features of the copulatory appara- layer of circular and longitudinal intermingled fibers tus, while the color pattern of Ch. marthae presents (10 µm thick). Vagina curving ventrally and opening two dark brown to black bands and a thin median onto female atrium (Figures 6A, 7). Female atrium light yellow band. Also, the pharynx is cylindrical. very short (Figures 6, 7, Table 3). Ratio between Regarding the copulatory apparatus, the course of length of female and male atrium between 18.9% and the ovovitelline ducts in Ch. marthae is very simi- 36.4% (Table 3). Female atrium lined by cuboidal lar to the new species, ascending posteriorly to the epithelium. Its muscularis formed by thin circular gonopore and joining dorsally a short common glan- layer (2.5 µm thick) and a longitudinal one (7–12 µm dular duct. The latter is approximately at the same thick). The latter adjacent to common muscle coat horizontal level as the prostatic vesicle, whereas in of variable thickness (62–187 µm thick), which is Ch. crassiphalla sp. nov. it is below the prostatic composed of longitudinal and oblique fibers. Scarce vesicle. Both in Ch. crassiphalla sp. nov. and Ch. erythrophil and cyanophil secretions open through marthae the distal portions of efferent ducts go the epithelium of the vagina and the female atrium through the bulbar musculature and ascend vertically (Figure 7B). to join the prostatic vesicle laterally. Nevertheless, Vitellaria well developed and abundant. In the pre- Ch. marthae has an almost spherical prostatic vesi- pharyngeal region, vitellaria extending both dorsally cle, smaller than in Ch. crassiphalla sp. nov., and and ventrally of intestine and between its branches this is anterior to the penis papilla. In addition, Ch.

Downloaded by [Duke University Libraries] at 08:39 12 December 2012 (Figure 4A). In the copulatory apparatus region, crassiphalla sp. nov. can be distinguished from Ch. vitellaria adjacent to ovovitelline ducts and common marthae because this species has a small and cylin- glandular duct (Figure 6A). drical penis papilla (Froehlich 1955), which is slightly Gonopore canal high, its epithelium cylindrical oblique and directed towards the posterior end of the and ciliated, receiving abundant cyanophil secretion body. and scarce erythrophil granulous secretion. Clearly, there is dissimilarity among the copulatory apparatus of the species of the genus Choeradoplana as some species have a permanent penis papilla while the Discussion others do not. Therefore, phylogenetic approaches are The features of the external and internal morphology needed to provide a better idea about the relationships of the new species ﬁt into Choeradoplana. The repro- of the Choeradoplana species. ductive system of this genus is highly heterogeneous, basically in regards to the copulatory apparatus. Except for Ch. ehrenreichi von Graff, 1899, for which Acknowledgments internal morphology has not yet been described, in The authors are grateful to Fundación Vida Silvestre Argentina Ch. banga Carbayo & Froehlich, 2012, Ch. bocaina and Ministerio de Ecología, Recursos Naturales Renovables y Studies on Neotropical Fauna and Environment 237

Turismo, Misiones for permission to conduct sampling at Reserva Froehlich CG. 1955. Sôbre morfologia e taxonomia das de Vida Silvestre Urugua-í, and especially to the park managers Geoplanidae. Bol Fac Fil Ciênc Letr, Sér Zool. 19:195–279. Ariel Tombo and Laura Aréjola for support during the fieldwork. Galindo-Leal C, Câmara G, editors. 2003. Atlantic forest hotspot We are thankful to the Administración de Parques Nacionales and status: an overview. In: The Atlantic Forest of South America: INTA San Antonio for permission to conduct sampling at the biodiversity status, threats, and outlook. Washington (DC): Campo Anexo, in San Antonio County. We wish to acknowledge Island Press. p. 3–11. Eudoxia M. Froehlich and Fernando Carbayo for sending us pho- Giraudo AR, Povedano H, Belgrano MJ, Krauczuk E, Pardiñas tos of the slides of the holotype of Choeradoplana marthae.The U, Miquelarena A, Ligier D, Baldo D, Castelino M. 2003. authors acknowledge the anonymous reviewer for their useful sug- Biodiversity status of the interior Atlantic forest of Argentina. In: gestions. This work was financed by CONICET (Consejo Nacional Galindo-Leal C, Câmara G, editors. The Atlantic Forest of South de Investigaciones Científicas y Técnicas), Agencia Nacional de America: biodiversity status, threats, and outlook. Washington Promoción Científica y Tecnológica (FONCyT) PICT 2007-01287, (DC): Island Press. p. 160–180. and UNLP (Universidad Nacional de La Plata) 11/N600. Marcus E. 1951. Turbellaria brasileiros (9). Bol Fac Fil Ciênc Letr, Sér Zool. 16:5–215. Negrete LHL, Brusa F, Winsor L. 2011. The blue land planarian References Caenoplana coerulea, an invader in Argentina. Rev Mex Biod. 82:287–291. Carbayo F, Froehlich EM. 2012. Three new Brazilian species of Ogren RE, Kawakatsu M. 1990. Index to the species of the the land planarian Choeradoplana (Platyhelminthes: Tricladida: family Geoplanidae (Turbellaria, Tricladida, Terricola) Part I: Geoplaninae), and an emendation of the genus. J Nat Hist. Geoplaninae. Bull Fuji Women’s Coll. 28(2):79–166. 46(19–20):1153–1177. Romeis B. 1989. Mikroskopische Technik. München (Germany): Carbayo F, Leal-Zanchet AM. 2003. Two new genera of geoplaninid Urban und Schwarzenberg. 697 p. land planarians (Platyhelminthes: Tricladida: Terricola) of Brazil von Graff L. 1894. Landplanarien. Viaggio del dott. Alfredo Borelli in the light of cephalic specializations. Inv Syst. 17(3):449–468. nella Republica Argentina e Paraguay, V. Boll Mus Zool Anat Di Bitetti MS, Placci G, Dietz LA. 2003. Una Visión de Comp Torino. 9(182):1–4. Biodiversidad para la Ecorregión del Bosque Atlántico del von Graff L. 1897. Neue Landplanarien. Viaggio del Dott. A. Alto Paraná: Diseño de un Paisaje para la Conservación de la Borelli nel Chaco Boliviano e nella Republica Argentina, IX. Boll Biodiversidad y prioridades para las acciones de conservación. Mus Zool Anat Comp Torino. 12(296):1–3. Washington (DC): World Wildlife Foundation. 154 p. von Graff L. 1899. Monographie der Turbellarien: II. Tricladida Du Bois-Reymond Marcus E. 1951. On South American Terricola (Landplanarien). Leipzig (Germany): Verlag von Geoplanids. Bol Fac Fil Ciênc Letr, Sér Zool. 16:217–255. Wilhelm Engelmann. 574 p. Downloaded by [Duke University Libraries] at 08:39 12 December 2012